Improved fluorescent compounds for tracing cell lineage

Abstract

In this note simple methods for the synthesis of several new fluorescent cell lineage tracers are described. These are fluorescent dextrans with average molecular weights of approximately 11 X 10(3), and with one or more fluorophore molecules covalently coupled to each dextran chain. These fluorescent dextrans are brighter than commercially obtainable products, and can be microinjected using either air-pressure injection or iontophoresis. They are long-lasting and have a uniform distribution in the cytoplasm of embryonic cells, clearly revealing very fine cell extensions such as cilia, axons, and filipodia. A method is also described for covalently attaching free amino groups to fluorescent dextran to make the tracers cofixable with cellular constituents by aldehyde treatment. Fluorescent dextran-amine tracers allow embryonic cell lineages to be studied in fixed, permeabilized, or sectioned embryos.